A Study on Developing the Planning Strategies of Smart City based on Smart Growth Concept

全球化浪潮及城市經年所積累之發展課題，將影響城市競爭力、魅力與吸引力之形塑，使政府須重新審視其一貫之發展與運作模式，以突破過往施政與規劃作為之盲點。於此情勢下，各國政府如何藉由靈活的規劃策略運用，以因應當前面臨之嚴峻挑戰，進而實踐理想之發展目標，著實為城市競爭力躍昇之關鍵。近年來許多國家為強化城市競爭力、找出自身發展契機，已著手智慧城市之研究與策略探討，使政府能針對弱勢發展面向研擬規劃策略，解決相關課題。 我國之智慧城市相關政策作為多聚焦於資訊通信能力之提升，較少重視其城市發展檢核功能之發揮，導致政府較難以察覺自身弱勢之所在；又，透過智慧城市目標之實踐能幫助其察覺自身發展優劣勢，以謀求改善發展問題之規劃策略面向。鑑於智慧城市之概念係源自智慧型成長理念而來，且二者皆有共同關注之城市發展議題。因此本研究企圖藉由智慧型成長理念建構之規劃策略應用，以促進城市健全發展與競爭力之提升。 爰此，本研究於文獻與理論分析之部分，先行歸納智慧型成長理念與智慧城市之意涵與應用面向，而後探討此二者間之關聯性，以釐清如何運用智慧型成長理念作為切中城市發展核心課題，進而建構達臻智慧城市各目標面向所需之智慧城市規劃策略。承上，本研究係以發展態樣多元之新北市為研究實證地區，並透過模糊德爾菲法(FDM)與分析網路程序法(ANP)兩階段問卷結果，篩選出新北市所需之智慧城市規劃策略，並排序其理想之施作優先順序。 最後，本研究以兩階段問卷結果為基礎，建立新北市智慧城市發展之架構，並研擬相關配套措施，以供未來政府推動智慧城市計畫之參考。而綜合上述之研究內容與成果，本研究可歸納三項結論，分別係：1.智慧型成長與智慧城市二者間應屬ㄧ動態之平衡關係；2.都市智慧發展課題之認知應係智慧城市推展之關鍵；3.智慧型成長理念建構之規劃策略有助智慧城市目標之達臻。同時，建議政府於智慧城市計畫推動之際，應增強各方對智慧城市意涵及其與智慧型成長二者關聯性之理解，動態調整規劃策略，成立相關政策推動之專責單位，賦予地方政府引領之角色，將行動方案法制化並確實落實規劃策略配套措施，以襄助城市能早日躋身智慧城市之列

Development of a Selective Medium and the Specific Primer for Detecting Ganoderma lucidum and Their Application

靈芝 (Ganoderma lucidum (Leyss. ex Fr.) Karst.) 具有高度的藥用價值，因而廣受菇農的栽種。為了要滿足市場的需求，故以太空包木屑培育靈芝後，會產生大量的廢棄太空包基質。這些廢棄的太空包基質常被用來製作成堆肥；惟若是堆肥中含有靈芝菌的活體時，則果樹會有被感染的風險。所以本研究嘗試研發選擇性培養基與聚合酶連鎖反應搭配專一性引子兩種方法，用以偵測堆肥中是否有存活的靈芝菌。靈芝菌選擇性培養基係利用 10 % V-8 培養基作為基礎基質，並添加 2 g l-1 腐絕 (40% ai)、200 mg l-1 滅達樂 (35% ai)、200 mg l-1 鏈黴素、0.2 g l-1 氯黴素、1 mg l-1 五氯硝基苯 (75% ai)、1.25 g l-1 單寧酸、10 ml l-1 酒精 (95% v/v) 及 2 ml l-1 乳酸 (85% v/v) 配製而成。靈芝菌選擇性培養基可由堆肥中分離出靈芝菌，且尚可抑制其他雜菌之生長。培養基中添加之單寧酸會使靈芝菌落周圍出現褐色反應，有助於靈芝菌的辨識。利用多重土丸取樣法分析不同靈芝接種源濃度之太空包木屑基質；結果顯示，靈芝菌選擇性培養基可偵測到台中及嘉義之太空包木屑基質經過 1000 倍無菌太空包基質稀釋後仍有 G. lucidum 的存活。此外，進一步研發聚合酶連鎖反應搭配專一性引子對，用以驗證靈芝菌選擇性培養基偵測靈芝菌之準確度。本研究由NCBI (National Center for Niotechnology information，USA.) 資料庫中篩選78株靈芝菌株之原始核苷酸 ITS1, 5.8S 及 ITS2 序列，設計出正向引子 GanJwF (5’-GCCTGCGTTTATCACAAACTC-3’) 搭配反向引子 ITS4-B (5’-CAGGAGACTTGTACACGGTCCAG-3’, Gardes and Bruns, 1993) 偵測靈芝菌，結果 GanJwF 與 ITS4-B 可增幅出約 600 bp 之條帶，且包含有大部分 ITS1, 5.8S 及 ITS2。該專一性引子對可偵測到含量僅 100 pg 之靈芝菌 DNA 且不與其他栽培種之擔子菌以及堆肥中之雜菌作用。結合這兩種偵測靈芝菌之方法可準確的偵測出廢棄太空包基質是否存活有靈芝菌。此外，利用熱及添加尿素的方式評估除滅靈芝菌之效果；結果顯示，將堆肥加熱至 55 ℃ 處理 10 分鐘或是添加 0.5 % 之尿素搭配 0.1 % 之生石灰處理 14 天，均可有效地將堆肥中之靈芝菌除滅。Ling-Chih, Ganoderma lucidum (Leyss. ex Fr.) Karst., is widely cultivated for its high medical value. In order to match the requirement of market, a large quantity of the sawdust is used for cultivation of Ling-Chih resulting in a lot of sawdust waste. The sawdust waste is often made as composts and used as fertilizers to supplement the nutrients for plants. However, if mycelia of Ling-Chih keep alive in composts, it may make high risk of fruit crops infected. Ganoderma spp. are the causal agents of basal stem rot (BSR) and mostly attack the root systems of the plants. In general, Ling-Chih is considered to be one of white-rot fungi and it is able to digest lignin, mainly. Large amounts of wood decay have an adverse effect on strength and stability, and make plants gradually weaken to die. Owing to the visible disease symptoms appear at a very late stage of infection, two detection methods, the selective medium and polymerase chain reaction (PCR) method based on taxon specific primer, were developed in this study to detect Ganoderma lucidum at early stage from compost. GLS medium was developed by amendment of 1 l 10% V-8 vegetable juice agar with 2 g thiabendazole (40% ai), 200 mg metalaxyl (35% ai), 200 mg streptomycin sulfate, 0.2 g chloramphenicol, 1 mg PCNB (75% ai), 1.25 g tannic acid, 10 ml ethanol (95% v/v), and 2 ml lactic acid (85% v/v). The GLS medium was able to rapidly isolate G. lucidum from the compost. Amendment of the medium with tannic acid could induce G. lucidum to produce brown halo surrounding the colony and make us easily recognize the fungus. To evaluate the efficacy of GLS medium, multiple-pellet soil-sampler method was used to assay the inoculum level of G. lucidum. The GLS medium was able to detect G. lucidum in the sawdust substrate from Taichung and Chiayi at 1000-fold dilution with sterilized sawdust substrate. The PCR method based on taxon specific primer was developed to confirm the detection results of GLS medium. The original nucleotide sequences of ITS1, 5.8S, and ITS2 of 78 isolates of G. lucidum were selected from NCBI GenBank (National Center for Biotechnology Information, USA). Based on the alignment results of ITS sequences, the forward primer GanJwF (5'-GCCTGCGTT-TATCACAAACTC-3') was designed and used with the reverse primer ITS4-B (5'-CAGG-AGACTTGTACACGGTCCAG-3', Gardes and Bruns, 1993) to detect G. lucidum. Approximate 600 bp PCR products containing most of the ITS1, 5.8S, and ITS2 regions could be amplified by primer pair of GanJwF and ITS4-B. PCR method based on GanJwF/ITS4-B specific primer pair is able to detect as low as 100 pg of G. lucidum DNA template and specific to G. lucidum, but not to other cultivated basidiomycetous or saprophytic fungi occurring in the compost. Two methods could used for for detecting the survival of G. lucidum in the compost and therefore be recommended for the manufacturer to determine if the compost is suitable as fertilizer. In order to eliminate G. lucidum from the compost, heat and urea treatments were used to suppress the growth of G. lucidum. The results showed that compost treated with 55 ℃ for 10 min or treated with 0.5% urea and 0.1% CaO for 10 days could completely eradicate the survival of G. lucidum.目次 中文摘要 ⅰ 英文摘要 ⅲ 目次 ⅴ 表次索引 ⅷ 圖次索引 ⅸ 前言 1 材料方法 6 一、供試靈芝菌株來源 6 二、供試菌株之培養與保存 6 三、溫度對靈芝菌絲生長之影響 6 四、供試菌株之鑑定 7 (一) DNA 萃取 7 (二) 聚合酶連鎖反應 7 (三) 靈芝 ITS 片段之電泳分析 8 (四) DNA序列解序與序列比對分析 8 五、研發選擇性培養基 9 (一) 基礎培養基篩選 9 (二) 酸鹼度對靈芝菌絲生長之影響 9 (三) 供試藥劑種類與篩選 10 (四) 酒精與乳酸對靈芝菌的影響 10 (五) GLS 培養基分離靈芝菌之靈敏度評估 11 (六) GLS 培養基抑菌族譜分析 11 1. 菌株來源 11 2. GLS 培養基的抑菌測試 12 六、靈芝菌專一性引子之開發與 PCR 測試 12 (一) 標準菌株之 ITS 基因序列擷取 12 (二) 靈芝 DNA 序列比對與專一性區域尋找 13 (三) 專一性引子設計 13 (四) 引子專一性之初步測試 13 (五) 不同黏合溫度對專一性引子對 GanJwF/ITS4-B 的影響 14 (六) 不同 Mg2+ 對專一性引子對 GanJwF/ITS4-B 的影響 15 (七) 檢測 GanJwF/ITS4-B 引子對對不同靈芝屬菌株之專一性 15 (八) 以 GanJwF/ITS4-B 引子對測試不同含量 DNA 之效果 15 (九) 專一性引子對對其他真菌之增幅效果 16 七、除滅靈芝菌之方法 16 (一) 致死溫度測試 17 (二) 醋酸與氨水對靈芝菌的抑制效果 17 (三) 不同尿素濃度對靈芝菌存活的影響 17 八、以 GLS 培養基與 GanJwF/ITS4-B 引子對之結合應用 18 結果 19 一、供試菌株之鑑定 19 二、溫度對靈芝菌絲生長之影響 19 三、靈芝選擇性培養基的研發 19 (一) 酸鹼度對靈芝菌絲生長之影響 19 (二) GLS 培養基的調製與其組成分對於靈芝菌生長的影響 20 (三) GLS 培養基分離靈芝菌之靈敏度評估 20 (四) GLS 培養基抑菌族譜分析結果 21 四、靈芝菌專一性引子之開發與 PCR 測試 22 (一) 專一性引子設計 22 (二) 引子專一性之初步測試 22 (三) 不同黏合溫度對專一性引子對 GanJwF/ITS4-B 的影響 23 (四) 不同 Mg2+ 濃度對於專一性引子對 GanJwF/ITS4-B 的影響 23 (五) 檢測 GanJwF/ITS4-B 引子對對不同靈芝菌株之專一性 23 (六) GanJwF/ITS4 引子對測試不同 DNA 濃度之效果 24 (七) 專一性引子對對其他真菌之增幅效果 24 五、除滅靈芝菌之方法 25 (一) 致死溫度測試 25 (二) 醋酸與氨水對靈芝菌的抑制效果 25 (三)不同尿素濃度對靈芝菌存活的影響 25 六、結合GLS 培養基與 GanJwF/ITS4-B 引子對之應用技術 26 討論 27 參考文獻 33 圖表 39 附錄 7

Development of a Selective Medium for Detecting Ganoderma lucidum in Compost

本研究主要目的在於研發靈芝菌的選擇性培養基，以便偵測廢棄的蕈菌生長基質調製成之堆肥或介質中是否仍有靈芝菌的存活。西元2009年，由嘉義取得的靈芝［Ganoderma lucidum （Leyss. ex Fr.）Karst.］太空包基質中分離GL1與GL3兩株靈芝菌菌株，進而評估七種真菌培養基、十種殺菌劑及兩種抗生素等對兩靈芝菌菌絲生長的影響，藉以研發靈芝菌的選擇性培養基。靈芝菌選擇性培養基（GLS）係利用10% V-8培養基作為基礎基質，並添加2 gL^(-1)腐絕（40% ai）、200 mg L^(-1)滅達樂（35% ai）、200 mg L^(-1)鏈黴素、200 mg L^(-1)氯黴素、1 mg L^(-1)五氯硝基苯（75% ai）、1.25 g L^(-1)單寧酸、10 mlL^(-1)酒精（95% v/v）及2 ml L^(-1)乳酸（85% v/v）配製而成。靈芝菌在GLS培養基生長2-3天可看到白色菌落，且於菌落周圍出現褐色反應。利用多重土丸取樣法分析含有不同靈芝接種源濃度之太空包木屑基質，結果顯示由台中及嘉義取得之靈芝太空包基質以無菌之太空包基質稀釋1000倍後，GLS培養基仍可偵測到G. lucidum的存活。本研究證明GLS培養基，除可自廢棄已久的太空包基質成功分離到靈芝菌外，尚可有效抑制其他雜菌的生長。 The purpose of this study was to develop a selective medium for detecting Ling-Chih, Ganoderma lucidum (Leyss. ex Fr.) Karst. in compost. In 2009, two isolates GL1 and GL3 of G. lucidum acquired from a Chiayi mushroom cultivation field were used to conduct the research. GLS (Ganoderma lucidum selective) medium was developed by amendment of 10% V-8 vegetable juice agar (1 L) with 2 g thiabendazole (40% ai), 200 mg metalaxyl (35% ai), 200 mg streptomycin sulfate, 200 mg chloramphenicol, 1 mg PCNB (75% ai), 1.25 g tannic acid, 10 ml ethanol (95% v/v), and 2 ml lactic acid (85% v/v). The GLS medium rapidly detected G. lucidum in the compost within 2-3 days. Amendment of the medium with tannic acid induced G. lucidum to produce brown halo surrounding the colony as a selective feature. The multiple-pellet soil-sampler method was used to evaluate the sensitivity of GLS medium. The GLS medium was able to detect G. lucidum in the spent sawdust substrate which were 1000-fold diluted in disinfected sawdust substrate. This study concluded that the GLS medium could detect the survival of G. lucidum in the compost, and inhibit the growth of other fungi. Thus, GLS medium was recommended for manufacturers to determine if the compost is a suitable fertilizer

研發偵測堆肥中靈芝菌的選擇性培養基

The purpose of this study was to develop a selective medium for detecting Ling-Chih, Ganoderma lucidum (Leyss. ex Fr.) Karst. in compost. In 2009, two isolates GL1 and GL3 of G. lucidum acquired from a Chiayi mushroom cultivation field were used to conduct the research. GLS (Ganoderma lucidum selective) medium was developed by amendment of 10% V-8 vegetable juice agar (1 L) with 2 g thiabendazole (40% ai), 200 mg metalaxyl (35% ai), 200 mg streptomycin sulfate, 200 mg chloramphenicol, 1 mg PCNB (75% ai), 1.25 g tannic acid, 10 ml ethanol (95% v/v), and 2 ml lactic acid (85% v/v). The GLS medium rapidly detected G. lucidum in the compost within 2-3 days. Amendment of the medium with tannic acid induced G. lucidum to produce brown halo surrounding the colony as a selective feature. The multiple-pellet soil-sampler method was used to evaluate the sensitivity of GLS medium. The GLS medium was able to detect G. lucidum in the spent sawdust substrate which were 1000-fold diluted in disinfected sawdust substrate. This study concluded that the GLS medium could detect the survival of G. lucidum in the compost, and inhibit the growth of other fungi. Thus, GLS medium was recommended for manufacturers to determine if the compost is a suitable fertilizer.本研究主要目的在於研發靈芝菌的選擇性培養基，以便偵測廢棄的蕈菌生長基質調製成之堆肥或介質中是否仍有靈芝菌的存活。西元2009年，由嘉義取得的靈芝［Ganoderma lucidum （Leyss. ex Fr.）Karst.］太空包基質中分離GL1與GL3兩株靈芝菌菌株，進而評估七種真菌培養基、十種殺菌劑及兩種抗生素等對兩靈芝菌菌絲生長的影響，藉以研發靈芝菌的選擇性培養基。靈芝菌選擇性培養基（GLS）係利用10% V-8培養基作為基礎基質，並添加2 gL^(-1)腐絕（40% ai）、200 mg L^(-1)滅達樂（35% ai）、200 mg L^(-1)鏈黴素、200 mg L^(-1)氯黴素、1 mg L^(-1)五氯硝基苯（75% ai）、1.25 g L^(-1)單寧酸、10 mlL^(-1)酒精（95% v/v）及2 ml L^(-1)乳酸（85% v/v）配製而成。靈芝菌在GLS培養基生長2-3天可看到白色菌落，且於菌落周圍出現褐色反應。利用多重土丸取樣法分析含有不同靈芝接種源濃度之太空包木屑基質，結果顯示由台中及嘉義取得之靈芝太空包基質以無菌之太空包基質稀釋1000倍後，GLS培養基仍可偵測到G. lucidum的存活。本研究證明GLS培養基，除可自廢棄已久的太空包基質成功分離到靈芝菌外，尚可有效抑制其他雜菌的生長